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Dept. Of Antitoxin King Institute of Preventive Medicine

Dept. Of Antitoxin King Institute of Preventive Medicine. Snake Venom Anti Sera against Naja naja Vipera russelli Bungarus caeruleus Echis carinatus . Snake Antivenom Sera for. Naja naja, Vipera russelli, Bungarus caeruleus, Echis carinatus. Snake Venom.

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Dept. Of Antitoxin King Institute of Preventive Medicine

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  1. Dept. Of Antitoxin King Institute of Preventive Medicine Snake Venom Anti Sera against Naja naja Vipera russelli Bungarus caeruleus Echis carinatus

  2. Snake Antivenom Sera for Naja naja,Vipera russelli, Bungarus caeruleus,Echis carinatus

  3. Snake Venom • More than 20 different constituents • Mainly proteins, enzymes and polypeptide toxins • Procoagulation enzymes • Haemorrhagins • Cytolytic or necrotic toxins • Haemolytic and myolytic phopholipases A2 • Presynaptic neurotoxins -Kraits • Post synaptic toxins - Cobra

  4. Preparation of the Snake Venom Solutions

  5. Immunization of the Equines with the Snake venom by the Standard Immunization Protocol

  6. Bleeding and separation of Plasma Test Bleeding , Plasma separated, RBC ‘s Reinfused

  7. PRODUCTION OF ANTIVENOM SERA: Technology in the public domain (not protected by intelectual property) A - Collection of venoms B – Horse Immunization Protocols C – Starting material of animal derived sera D – Fractionation & Purification process

  8. Pepsin digestion of IgG. Digestion normally produces a F(ab´)2 fragment and an extensively degraded Fc portion.

  9. TYPICAL FRACTIONATION PROCESS OF F(ab’)2 FRAGMENTS Hyperimmune plasma Acidification at pH 3.0-3.3, digestion with pepsin at 30°C for 1 hr Addition of 12% ammonium sulphate Heating to 56°C for 1 hr with stirring Adjust pH to 5.2 Filtration or centrifugation Discard the precipitate Filtrate or supernatant Addition of ammonium sulphate to reach 23% concentration; stir for 1 hr Filtration or centrifugation Discard the supernatant F(ab̒)2 solution

  10. Solubilization of precipitate. Tangential flow filtration and concentration Concentrated F(ab̒)2 solution sterile filtration Final Product

  11. Salient features of ASVS production Long ,laborious and time consuming, process (about 1 year) Maintenance of costly and sensitive animals -Horses and mules Increasing demand for the specific life saving drug by the TN state , highly necessary to augument the production of AntiSnake Venom serum. The annual requirement of ASVS serum is about 5 lakh vials 1 vial -10 ml

  12. King Institute is manufacturing ASVS since the year 1980 and has supplied to all the Govt. hospitals in the state of Tamil naduThe maximum number of vials was produced during year 1997-1998 of 94,458 vials • 2000-2001 30194 • 2001-2002 4304 • 2002-2004 Nil • 2004-2005 2340 • 2005-2006 Nil • 2006-2007 3161 • 2007-2008 Nil • 2008-2009 Nil Last SUPPLIED -1000 vials during June 2012

  13. Important issues

  14. The Govt. of Tamil nadu has sanctioned an amount of Rs. 7.36 crores for the construction of New Building as per c GMP Norms as Phase I allotment. The requistion for Rs 12.14 crores is to be submitted as Phase II allotment for Equipmens,Horse purchase ,Civil and Eletrical Modifications ,Manpower and Venoms and chemicals .

  15. Assessment of neutralizing dose of RV by monovalent antiserum by ELISA • Production of IgY Antibodies against RV. • Production of bivalent antibodies against RV+EV

  16. Thank you

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